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Figure 3 | Cancer & Metabolism

Figure 3

From: An epitope tag alters phosphoglycerate dehydrogenase structure and impairs ability to support cell proliferation

Figure 3

Purified untagged recombinant PHGDH has higher specific activity than N-terminally tagged PHGDH. (A) Coomassie gel of rPHGDH (untagged) after each step of the purification: 1. cleared lysate from E. coli expressing rPHGDH protein, 2. eluate from Cibacron Blue F3GA column, 3. eluate from anion exchange column, 4. eluate from size exclusion column, and 5. rFLAG-His-PHGDH purified by sequential nickel and size exclusion columns, for comparison. Molecular weight standards in kilodaltons are present to the left of lane 1. (B) In vitro enzyme activity of untagged rPHGDH. Assay was performed at saturating substrate concentrations. (C) Comparison of relative specific activity of rPHGDH and rHis-PHGDH. Error bars show standard deviation from the mean.

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Cancer & Metabolism

ISSN: 2049-3002